Antiserum for Insulin Radioimmunoassay Generated by an Insulin Derivative Devoid of Hypoglycemic Activity

Abstract

A tetraiodinated derivative of bovine insulin, prepared at pH 1 with stable iodine, was unable to cause signs of hypoglycemia in doses up to 2.4 micrograms/g in fasting mice, when native insulin caused 100% mortality. In neutral and acidic solutions, in absence of chaotropic agents, it behaved as the monomer, and could be separated from less iodinated, active species, that appeared as dimers, by conventional gel filtration. To generate antibodies in guinea-pigs, the tetraiodinated insulin was injected in doses three times higher than native insulin, without any harm to recipient animals. The induced antiserum was compared with antiserum generated by conventional methods in radioimmunoassay (RIA) of native insulin, and parallel curves were obtained.